A Panel of Real-Time PCR Assays for the Direct Detection of All of the Xylella fastidiosa Subspecies

The bacterial plant pathogen Xylella fastidiosa causes disease in hundreds of plant species worldwide including many crops, and as such accurate determination of the subspecies of the bacteria is vital to control, containment, and eradication measures. Conventional methods to determine the subspecies of X. fastidiosa rely on time consuming multilocus sequence typing (MLST), a laborious multistage process. This chapter provides a rapid alternative to MLST utilizing real-time PCR assays to provide highly specific and sensitive detection of the pathogen subspecies. Here we describe the methodology for sampling plant material, performing the DNA extraction and undertaking the real-time PCR assays. This method allows straightforward, robust, reliable, high-throughput, and rapid determination of the X. fastidiosa subspecies.